Smad4 | HUABIO HA750070 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

Smad4

catalog :

HA750070

quantity :

100μl

price :

649.00 USD

clonality :

monoclonal

host :

domestic rabbit

conjugate :

nonconjugated

clone name :

SP05-05

reactivity :

human, mouse, rat

application :

western blot, flow cytometry, immunohistochemistry - paraffin section

more info or order :

HUABIO product webpage

image

image 1 :

Western blot analysis of Smad4 on different lysates with Rabbit anti-Smad4 antibody (HA750070) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HCT 116 cell lysate Lane 3: C6 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 60 kDa Observed band size: 70 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750070) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

image 2 :

Western blot analysis of Smad4 on Neuro-2a cell lysates with Rabbit anti-Smad4 antibody (HA750070) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 60 kDa Observed band size: 70 kDa Exposure time: 3 minutes; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750070) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

image 3 :

All lanes: Western blot analysis of Smad4 with anti-Smad4 antibody (HA750070) at 1:1,000 dilution. Lane 1: Wild-type HaCaT whole cell lysate (15 µg). Lane 2: Smad4 knockout HaCaT whole cell lysate (15 µg). ET1604-12 was shown to specifically react with Smad4 in wild-type HaCaT cells. NO band was observed when Smad4 knockout sample was tested. Wild-type and Smad4 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1604-12, 1:1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody at 1:10,000 dilution was used for 1 hour at room temperature.

product information

SKU :

HA750070

Target name :

Smad4

Species reactivity :

Human,Mouse,Rat

Applications :

WB,IF-Cell,IF-Tissue,IHC-P,FC

Conjugate :

Non-conjugated

Immunogen :

Synthetic peptide within C-terminal human Smad4.

Uniprot id :

Q13485>SwissProt: Q13485 Human;SwissProt: P97471 Mouse;SwissProt: O70437 Rat

Host :

Rabbit

Clone number :

SP05-05

Isotype :

IgG

Size :

100μl

List Price :

649.00 USD

Storage Buffer :

PBS (pH7.4).

Form :

Liquid

Storage Instruction :

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Purity :

Protein A affinity purified.

Product type :

Recombinant Rabbit monoclonal Antibody

Positive control :

HeLa cell lysate, HCT 116 cell lysate, C6 cell lysate, Neuro-2a cell lysates, HeLa, Neuro-2a, PC-12, human colon tissue, human lung tissue, mouse lung tissue, rat lung tissue.

Molecular wt :

Predicted band size: 60 kDa

Subcellular location :

Cytoplasm, Nucleus.

Concentration :

1 mg/mL.

Recommended dilutions :

WB: 1:1,000-1:5,000 ;IF-Cell: 1:100 ;IF-Tissue: 1:50-1:200 ;IHC-P: 1:200 ;FC: 1:1,000

Advanced Validation :

Knockout (KO)

Pic img4 :

https://storage.huabio.cn/huabio/productImg/HA750070_4.jpg

Pic legend4 :

Immunocytochemistry analysis of HeLa cells labeling Smad4 with Rabbit anti-Smad4 antibody (HA750070) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Smad4 antibody (HA750070) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Pic img5 :

https://storage.huabio.cn/huabio/productImg/HA750070_5.jpg

Pic legend5 :

Immunocytochemistry analysis of Neuro-2a cells labeling Smad4 with Rabbit anti-Smad4 antibody (HA750070) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Smad4 antibody (HA750070) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Pic img6 :

https://storage.huabio.cn/huabio/productImg/HA750070_6.jpg

Pic legend6 :

Immunocytochemistry analysis of PC-12 cells labeling Smad4 with Rabbit anti-Smad4 antibody (HA750070) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Smad4 antibody (HA750070) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Pic img7 :

https://storage.huabio.cn/huabio/productImg/HA750070_7.jpg

Pic legend7 :

Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Smad4 antibody (HA750070) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750070) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img8 :

https://storage.huabio.cn/huabio/productImg/HA750070_8.jpg

Pic legend8 :

Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-Smad4 antibody (HA750070) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750070) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img9 :

https://storage.huabio.cn/huabio/productImg/HA750070_9.jpg

Pic legend9 :

Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Smad4 antibody (HA750070) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750070) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img10 :

https://storage.huabio.cn/huabio/productImg/HA750070_10.jpg

Pic legend10 :

Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-Smad4 antibody (HA750070) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750070) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Pic img11 :

https://storage.huabio.cn/huabio/productImg/HA750070_11.jpg

Pic legend11 :

Flow cytometric analysis of HeLa cells labeling Smad4. Cells were fixed and permeabilized. Then stained with the primary antibody (HA750070, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Pic img12 :

https://storage.huabio.cn/huabio/productImg/HA750070_12.jpg

Pic legend12 :

Flow cytometric analysis of Neuro-2a cells labeling Smad4. Cells were fixed and permeabilized. Then stained with the primary antibody (HA750070, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Pic img13 :

https://storage.huabio.cn/huabio/productImg/HA750070_13.jpg

Pic legend13 :

Flow cytometric analysis of PC-12 cells labeling Smad4. Cells were fixed and permeabilized. Then stained with the primary antibody (HA750070, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

more info or order :

HUABIO product webpage